Tez (Yüksek Lisans) - Süleyman Demirel Üniversitesi, Sağlık Bilimleri Enstitüsü, Anatomi Anabilim Dalı, 2002. / Bibliyografya var.

The excitation-contraction-relaxation cycle of the guinea pig ileum longitudinal smooth muscle was studied in muscles contracted by a muscarinic agent, cis-2-methyl-4-dimethylaminomethyl-1,3-dioxolane methiodide (CD) and by 60 mM KC1. Aspects of the cycle were investigated by analyzing the active transport enzyme activities in the sarcolemma, the tissue Ca depots which could release Ca for contraction and the sensitivity of the contractile responses to extracellular ion changes. Essentially net changes of intracellular Ca, Mg, Na and K content during contractions were measured by a modified 'La method'. The tissues were washed for 30 min in 160 mM Tris-HCl solution (pH 7.4) containing 10 mM LaCl₃ at 4°C in order to seal the intracellular ions in the cell and displace extracellular ions. A method to loosen the 'intercellular cementing' substance by reducing the tissue Ca and Mg was developed as an adjunct to the preparation of a sarcolemmal enriched microsomal fraction. The method reduced the tenacity of the tissue and made the tissue easy to disrupt by a mild homogenizing procedure. The method also appeared to aid the extraction of contractile proteins. The microsomal fraction was not detectably contaminated by mitochondria and was enriched with vesicles of sarcolemma, probably originating from the muscle caveolae. The sarcolemma enriched microsomal fraction had a Ca-ATPase activity that was progressively stimulated by 10⁻⁷ to 2.4 x 10⁻⁴ M free Ca²⁺ , did not require Mg and was inhibited by La. The microsomal Ca-ATPase activity was not due to contamination by actomyosin. The actomyosin Ca-r-ATPase in the soluble fraction had a higher affinity than the microsomal Ca-ATPase for Ca and for La. The microsomal Ca-ATPase activity was postulated to be associated with an active Ca pump thought to he located in the cayeolae. The microsomal fraction had a Mg-dependent ATPase that could Be stimulated by Na, but K and ouabain had very little additional effect. The addition of an activating factor in the soluble fraction conferred some K and ouabain sensitivity to the Mg-dependent Na-ATPase, which indicated that a Na,K-ATPase was present in this tissue. Low doses of ouabain contracted the longitudinal ileum but the responses were not antagonized by raising the external K concentration five fold, as would be expected if ouabain acted by inhibiting the Na,K-ATPase. However, the ouabain response was rapidly lost when extracellular Ca was removed from the medium and the decline of the response followed the same time course as the loss of extracellular Ca. The peak of the ouabain contraction coincided with significant increases of intracellular Ca and Na, but K loss was not apparent until relaxation ensued. The results suggested that ouabain has an early direct effect on membrane permeability before it inhibited the Na,K-ATPase. CD (2 x 10⁻⁷ M) and 60 mM KCl induced phasic and tonic contractions of the longitudinal muscle of the ileum. The phasic contraction declined from 100% to 7% over 10 min when Ca was omitted from the physiological medium. This decline followed the time course of the loss of extracellular Ca. This, together with the fact that low concentrations of LaCl₃ inhibited the phasic component, indicated that Ca bound to the outer aspect of the cell was responsible for the phasic component. The tonic component was lost more rapidly than the phasic component when the Ca was removed from the Tyrode's solution. The tonic component seemed activated by free Ca mobilized from the extracellular space. The extracellular origin of the Ca for contraction was consistent with the observed small net gain of intracellular Ca that occurred during the phasic and tonic contractions. The minimal volume of the sarcoplasmic reticulum and the abundance of caveolae was also consistent with the high sensitivity of the tissue to extracellular Ca concentrations. The intracellular Ca gained during contraction wa,s extruded within 30 sec after the CD or 60 mM KCl were washed out of the tissue bath, Following washout of CD, the muscle was quiescent for the 20 to 30 min 'equilibration' phase. Spontaneous activity was absent during this phase and tension was below baseline. After a maximal CD contraction, a second response to CD or to 60 mM KCl induced during the 'equilibration' phase had an altered or desensitized biphasic appearance. Responses of the muscle to CD for 10 min were accompanied by a cytoplasmic loss of K. After washout of CD, the K was regained slowly over 20 to 30 min. Stimulation of the tissue by 60 mM KCl did not cause a loss of K from the muscle nor did it cause desensitization of the muscle. Higher extracellular K concentrations decreased the time required after CD contractions for the return of spontaneous activity and prevented muscle desensitization to repeated doses of CD, probably by accelerating the return of intracellular K levels to normal. It was proposed that during contraction, elevated intracellular Ca activated K channels, thereby increasing K permeability and causing the 'after-hyperpolarization' and subsequent desensitization which follows muscarinic induced contractions. / Pharmaceutical Sciences, Faculty of / Graduate

A distrofia muscular do Golden Retriever é uma miopatia hereditária, recessiva e fatal. Achados clinicopatológicos no trato gastrintestinal desses cães, como atrofia muscular, megaesôfago, dilatação gástrica são relatados, com possíveis alterações nos plexos entéricos. Este trabalho tem como objetivo analisar os neurônios colinérgicos e nitrérgicos do plexo mioentérico, a expressão do receptor P2X7 e a morfologia do íleo de cães afetados pela distrofia muscular comparados aos de cães não afetados. Os tecidos foram preparados por métodos imunohistoquímicos de marcação do Óxido Nítrico Sintase (NOS), Acetilcolina Transferase (ChAT), do pan-neuronal anti-HuC/D e do receptor P2X7. As análises qualitativas e quantitativas das contagens das marcações, das densidades neuronais e da área dos perfis foram obtidas dos Microscópios de Fluorescência, de Confocal de Varredura à Laser, e Microscópio Eletrônico de Transmissão. Os resultados qualitativos demonstraram que neurônios NOS-ir e ChAT-ir apresentaram morfologia Dogiel Tipo I com fibras que colocalizam com o receptor P2X7 e a musculatura intestinal com núcleos picnóticos e maior quantidade de fibras colágenas no grupo distrófico. Os dados quantitativos demonstraram: a) diminuição na área do perfil neuronal dos neurônios NOS-ir e ChAT-ir no grupo distrófico b) maior densidade de neurônios NOS-ir no grupo distrófico. O presente estudo adicionou informações sobre o código químico do plexo mioentérico de cães que podem facilitar o entendimento de desordens intestinais nesses animais. / The Golden Retriever muscular dystrophy is a hereditary myopathy, recessive and fatal. Clinical and pathological findings in the gastrointestinal tract of these dogs as muscle atrophy, megaesophagus, and gastric dilatation are reported, with possible changes in the enteric plexus. This work aims to analyze the cholinergic and nitrergic neurons of myenteric plexus, the P2X7 receptor expression and morphology of the ileum of dogs affected by muscular dystrophy compared to those of unaffected dogs. Tissues were prepared by immunohistochemical methods of labeling Nitric Oxide Synthase (NOS), acetylcholine transferase (ChAT), the panneuronal anti-HuC / D and P2X7 receptor. Qualitative and quantitative analyzes of scores of labeling, density and of neuronal profiles area were obtained from Fluorescence Microscopes, Confocal Laser Scanning, and Transmission Electron Microscope. The results showed that NOS-(immunoreactive)ir and ChAT-ir neurons present morphology Dogiel Type I, their fibers colocalize with the P2X7 receptor and the intestinal muscles present pyknotic nuclei with increased amount of collagen fibers in the dystrophic group. The quantitative data showed: a) decrease in the neuronal area profile of NOS-ir and ChAT-ir neurons in the dystrophic group b) higher density of NOS-ir neurons in the dystrophic group. The present study added information about the chemical code of the myenteric plexus of dogs that can facilitate the understanding of intestinal disorders in these animals.

Diss. (sammanfattning) Uppsala : Uppsala universitet, 2007. / Härtill 4 uppsatser.

Submitted by Rafael Santana (rafael.silvasantana@ufpe.br) on 2017-10-31T18:35:50Z No. of bitstreams: 2 license_rdf: 811 bytes, checksum: e39d27027a6cc9cb039ad269a5db8e34 (MD5) DISSERTAÇÃO MARJORIE 1.pdf: 1501690 bytes, checksum: cff8cd81b84e5a52cfea3b2b854df884 (MD5) / Made available in DSpace on 2017-10-31T18:35:50Z (GMT). No. of bitstreams: 2 license_rdf: 811 bytes, checksum: e39d27027a6cc9cb039ad269a5db8e34 (MD5) DISSERTAÇÃO MARJORIE 1.pdf: 1501690 bytes, checksum: cff8cd81b84e5a52cfea3b2b854df884 (MD5) Previous issue date: 2015-11-27 / FACEPE / Simarouba amara (Simaroubaceae) é popularmente conhecida como "pau-paraíba" ou “marupá”. No Brasil, é encontrada em todo o litoral brasileiro e tem sido usada na medicina tradicional, principalmente no tratamento de alterações de origens inflamatória, gástrica e respiratória. Esse estudo avaliou a possível atividade espasmolítica do extrato aquoso da casca do caule de Simarouba amara (EASa). Nesse sentido, ensaios in vitro de curvas concentração-efeito (CCE) simples ou cumulativas foram obtidas para histamina, carbacol, KCl e CaCl2 na ausência ou presença de EASa ou verapamil (VRP) no íleo isolado de cobaia. Os resultados mostram que o EASa (125, 250 e 500 µg/mL) e o VRP (0,025 a 3,2 µM) reduziram estatisticamente os componentes fásicos e tônicos das CCE simples para histamina, carbacol e KCl. Nas CCE cumulativas induzidas por histamina e carbacol, o ESSa e VRP produziram redução significativa do efeito máximo (Emáx) sem deslocamento das curvas à direita, características de antagonismo do tipo não competitivo. Nas curvas cumulativas de CaCl2, EASa e VRP reduziram significativamente o Emáx, contudo com deslocamento da curva à direita, que no caso do EASa foi observado apenas com a maior dose. Na presença do anti-histamínico H1 hidroxizina (HXZ, 0,125 e 0,250 μM), observou-se redução do Emáx das CCE cumulativas de histamina de 100,00 ± 0,00 para 84,53 ± 6,44% e 63,25 ± 4,63% respectivamente e deslocamento significativo à direita (pCE50 controle = 7,08 ± 0,04 para pCE50 = 5,46 ± 0,28 e 5,54 ± 0,19) respectivamente. Quando o EASa (250 e 500 µg/mL) foi associado ao HXZ houve potencialização significativa da redução do Emáx 32,72 ± 1,95% e 17,83 ± 2,48% respectivamente, todavia, os deslocamentos das curvas cumulativas à direita (pCE50 = 5,13 ± 0,23 e 4,80 ± 0,74) não foram estatisticamente diferentes das obtidas na presença de hidroxizina. O conjunto dos resultados sugere possível ação espasmolítica não seletiva do extrato aquoso da casca de Simarouba amara, a qual foi correlacionada inicialmente ao bloqueio do influxo de cálcio através dos canais de cálcio voltagem dependentes. / Simarouba amara (Simaroubaceae) is popularly known as "pau-paraíba" or "marupá". In Brazil, it is found throughout the Brazilian coast and has been used in traditional medicine, especially in the treatment of disorders of inflammatory origins, gastric and respiratory. This study evaluated the possible spasmolytic activity of the aqueous extract of the bark of Simarouba amara . Accordingly, in vitro concentrationeffect curves (CCE) were obtained from single or cumulative histamine, carbachol, KCl and CaCl2 in the absence or presence of this or verapamil on isolated guinea pig ileum. The results show that the EASa (125, 250 and 500 µg / ml), and VRP (3,2 0.025 mM) statistically reduced the phasic and tonic components of the single CCE histamine, carbachol and KCl. The cumulative CCE induced by histamine and carbachol, the VRP and this produced a significant reduction of the maximum effect (Emax) without shifting the curves to the right, antagonism characteristics of the noncompetitive type. In the cumulative curve CaCl2, and EASa VRP Emax significantly reduced, yet with displacement of the right turn, in which case the EASa was observed only at the highest dose. In the presence of H1 antihistamine hydroxyzine (HXZ, 0.125 and 0.250 mM), there was reduction of Emax pCE50 cumulative 100.00 ± 0.00 to 84.53 ± histamine 6.44% and 63.25 ± 4.63% respectively and significant shift to the right (pCE50 control = 7.08 ± 0.04 to 5.46 ± 0.28 = pCE50 and 5.54 ± 0.19) respectively. When the EASA (250 and 500 mg / mL) was associated with HXZ significant potentiation of the decrease Emax% 32.72 ± 1.95 and 17.83 ± 2.48%, respectively, however, the displacements of the curves to the right of cumulative (pCE50 = 5.13 ± 0.23 and 4.80 ± 0.74) were not statistically different from those obtained in the presence of hydroxyzine. The set of results suggests possible nonselective spasmolytic action of the aqueous extract of Simarouba loved bark, which was initially correlated to block the calcium influx through voltage gated calcium channels.

A distrofia muscular do Golden Retriever é uma miopatia hereditária, recessiva e fatal. Achados clinicopatológicos no trato gastrintestinal desses cães, como atrofia muscular, megaesôfago, dilatação gástrica são relatados, com possíveis alterações nos plexos entéricos. Este trabalho tem como objetivo analisar os neurônios colinérgicos e nitrérgicos do plexo mioentérico, a expressão do receptor P2X7 e a morfologia do íleo de cães afetados pela distrofia muscular comparados aos de cães não afetados. Os tecidos foram preparados por métodos imunohistoquímicos de marcação do Óxido Nítrico Sintase (NOS), Acetilcolina Transferase (ChAT), do pan-neuronal anti-HuC/D e do receptor P2X7. As análises qualitativas e quantitativas das contagens das marcações, das densidades neuronais e da área dos perfis foram obtidas dos Microscópios de Fluorescência, de Confocal de Varredura à Laser, e Microscópio Eletrônico de Transmissão. Os resultados qualitativos demonstraram que neurônios NOS-ir e ChAT-ir apresentaram morfologia Dogiel Tipo I com fibras que colocalizam com o receptor P2X7 e a musculatura intestinal com núcleos picnóticos e maior quantidade de fibras colágenas no grupo distrófico. Os dados quantitativos demonstraram: a) diminuição na área do perfil neuronal dos neurônios NOS-ir e ChAT-ir no grupo distrófico b) maior densidade de neurônios NOS-ir no grupo distrófico. O presente estudo adicionou informações sobre o código químico do plexo mioentérico de cães que podem facilitar o entendimento de desordens intestinais nesses animais. / The Golden Retriever muscular dystrophy is a hereditary myopathy, recessive and fatal. Clinical and pathological findings in the gastrointestinal tract of these dogs as muscle atrophy, megaesophagus, and gastric dilatation are reported, with possible changes in the enteric plexus. This work aims to analyze the cholinergic and nitrergic neurons of myenteric plexus, the P2X7 receptor expression and morphology of the ileum of dogs affected by muscular dystrophy compared to those of unaffected dogs. Tissues were prepared by immunohistochemical methods of labeling Nitric Oxide Synthase (NOS), acetylcholine transferase (ChAT), the panneuronal anti-HuC / D and P2X7 receptor. Qualitative and quantitative analyzes of scores of labeling, density and of neuronal profiles area were obtained from Fluorescence Microscopes, Confocal Laser Scanning, and Transmission Electron Microscope. The results showed that NOS-(immunoreactive)ir and ChAT-ir neurons present morphology Dogiel Type I, their fibers colocalize with the P2X7 receptor and the intestinal muscles present pyknotic nuclei with increased amount of collagen fibers in the dystrophic group. The quantitative data showed: a) decrease in the neuronal area profile of NOS-ir and ChAT-ir neurons in the dystrophic group b) higher density of NOS-ir neurons in the dystrophic group. The present study added information about the chemical code of the myenteric plexus of dogs that can facilitate the understanding of intestinal disorders in these animals.

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Evidências indicam uma interação na comunicação glia-neurônio. E sabido que a isquemia e reperfusão intestinal (I/R) são eventos clínicos graves. Este projeto visa estudar os efeitos de Brilliant Blue G (BBG) e de Probenecida (PB) nas células gliais entéricas (CGE). Os vasos ileais foram ocluídos por 45 min. Os períodos de reperfusão foram de 24 h, 14 e 28 dias.Os animais foram tratados com BBG, PB ou Salina. Os dados mostraram que o receptor P2X7 estava em CEG e neurônios, que há fenótipos diferentes de CGE e Panx-1 estava em CEG (GFAP). As densidades mostraram diminuição do número de células-IR ao receptor P2X7, Panx-1 e Hu no grupo I/R Salina, porém nos grupos tratados houve recuperação do número dessas células e com as CGE ocorreu um aumento no grupo I/R Salina. A área do perfil celular apresentou alterações em neurônios e CGE. Houve alterações na expressão proteica de P2X7 e Panx-1 e na atividade contrátil do intestino. O uso de BBG e de PB têm sido eficaz na recuperação de CGE e neurônios e podem ser alvos terapêuticos para doenças do trato gastrintestinal. / Evidence indicates an interaction in glia-neuron communication. It is known that intestinal ischemia and reperfusion (I/R) are serious clinical events. This project aims to study the effects of Brilliant Blue G (BBG) and Probenecida (PB) on enteric glial cells (CGE). The ileal vessels were occluded for 45 min. Reperfusion periods were 24 h, 14 and 28 days. Animals were treated with BBG, PB or Saline. The data showed that the P2X7 receptor was in CEG and neurons, that there are different phenotypes of CGE and Panx-1 was in CEG (GFAP). The densities showed a decrease in the number of cells-IR to the P2X7 receptor, Panx-1 and Hu in the Salina I/R group, but in the treated groups there was a recovery in the number of these cells and with the CGE there was an increase in the Salina I/R group. The cell profile area presented changes in neurons and CGE. There were changes in the protein expression of P2X7 and Panx-1 and in the contractile activity of the intestine. The use of BBG and PB has been effective in the recovery of CGE and neurons may be therapeutic targets for diseases of the gastrointestinal tract.